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.2020 Jan 14;9(1):58.
doi: 10.3390/pathogens9010058.

Evaluation of Hypoxia-Inducible Factor-1 Alpha (HIF-1α) in Equine Sarcoid: An Immunohistochemical and Biochemical Study

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Evaluation of Hypoxia-Inducible Factor-1 Alpha (HIF-1α) in Equine Sarcoid: An Immunohistochemical and Biochemical Study

Manuela Martano et al. Pathogens..

Abstract

Background: equine sarcoids are the most frequent skin tumors in equidae worldwide. It is well known that delta bovine papillomaviruses are their causative agents. We have recently shown the presence in equine sarcoids of abnormal vessel structures, which could cause a hypoxic condition. The aim of this study was to analyze the expression of hypoxia-inducible factor-1 alpha (HIF-1α) in a subset of BPV positive equine sarcoids and explore the relationship with vascular endothelial growth factor (VEGF) expression.

Results: 80% of equine sarcoids showed strong cytoplasmic staining in >60% of neoplastic fibroblasts, while 20% of samples showed a moderate cytoplasmic staining in 40-60% of neoplastic fibroblasts for HIF-1α. Results of Western blotting (WB) were consistent with immunohistochemistry (IHC). Moreover, a positive correlation between HIF-1α and VEGF expression (r = 0.60,p < 0.01) was observed.

Conclusion: we have shown that HIF-1α was strongly expressed in equine sarcoid. The upregulation of HIF-1α has been described in numerous tumors and can be modulated by many proteins encoded by transforming viruses. Thus, it is also possible that BPV could have a relevant role in HIF-1α pathway regulation, contributing to the development of equine sarcoids by promoting HIF-1α/VEGF mediated tumor angiogenesis.

Keywords: BPV; HIF-1α; VEGF; equine; sarcoid.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Equine normal skin. HIF-1α immunohistochemical staining. Expression in the basal epidermal cells (arrow) (40×). Scale bar: 100 μm.
Figure 2
Figure 2
Equine sarcoid. HIF-1α immunohistochemical staining. (a) Strong immunohistochemical expression in sarcoid fibroblasts (arrow) (score 3; ++; 40×; Scale bar: 100 μm). (b) Moderate immunohistochemical expression in sarcoid fibroblasts (arrow) (score 2; +; 20×; Scale bar: 100 μm).
Figure 3
Figure 3
(A) Western blotting analysis showing overexpression of HIF-1α in equine sarcoids (S) compared to normal skin samples (N). Whole cell lysate from Hela cells and K562 cells was run concomitantly to ensure the specificity of the band. Blot was stripped and incubated with anti-β-actin antibody to perform normalization. (B) Densitometric analysis was performed with the results expressed as HIF-1α/actin ratio.
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